P75在小鼠肾脏组织中的特异性表达鉴定

Identify the expression of P75 in the mouse kidney

目的探讨肾脏中是否存在神经嵴来源的细胞,体外分离后培养,检测其细胞特性。方法通过免疫荧光染色方法及Q-PCR技术,检测P75在小鼠肾脏中是否有表达及在各时间段(1、2、4周)的表达规律;用免疫荧光共染的方法,分别行P75抗体与Nestin抗体、Nephrin抗体共染,观察P75+细胞表达位置;磁珠分选P75+的细胞,并在体外培养及鉴定。结果P75在小鼠肾脏中有表达,在7 d时表达含量最高,达18.7%;随小鼠年龄的增长,P75表达含量降低;P75与Nestin、Nephrin的表达部位一致,在肾小球中表达,而未在肾小管表达;磁珠分选得到的P75+细胞经可体外培养;并在诱导液诱导下,这些细胞可以分化为脂肪细胞和成骨细胞。结论P75表达阳性的肾脏细胞,有可能起源自神经嵴,且具有部分干细胞特性。

Objective To investigate whether there is a group of cells which originates in the neural crest, isolate and culture the kind of cells in vitro for analysis itscharacteristics. Methods By immunofluorescence staining method and Q-PCR technology＇, we detected P75 expression in the mouse kidney and the expression pattern of each time period （one week, two weeks, four weeks） ; P75 antibody waseo-stained with Nestin and Nephrin antibody in the mouse kidney by immunofluorescenee method; P75-positive cells were sorted, cultured identifiedin vitro. Results P75 expression was detectedin the mouse kidney, andits levels was peaked at postnatal 7-day （about18.7%）; and the expression was gradually degreasedwith growth. Such P75-positive cells could be co-stained with Nestin and Nephrin antibody in the glomerulus. The P75-positive cells could be sorted about 15.3% and cultured in vitro, and could differentiate intoadipocytes and osteoblast. Conclusion P75-positive kidney cells maybe originate from the neural crest, and revealedcharacteristics of stem cells.